Search

Aims Belowground legume nitrogen (N) composed of roots and rhizodeposition is an important N input to soils, but published data of belowground N vary broadly, probably due to extrapolation from short-term experiments and dissimilar growing conditions. We quantified belowground N inputs of red clover (Trifolium pratense L.) during two consecutive years in a clover-grass sward along a soil nutrient availability gradient. Methods We established a red clover-perennial ryegrass (Lolium perenne L.) model sward in microplots located in field plots of the DOK experiment, which has a 33-year history of organic and conventional cropping, resulting in a soil nutrient availability gradient. Four treatments were examined: the zero fertilisation control, bio-organic with half and full dose manure application, and the conventional system with mineral fertilisation at full dose. We studied the development of clover aboveground and belowground N using multiple pulse 15N urea leaf labelling. Results Belowground clover N increased over time and with rising nutrient availability and was proportional to aboveground clover N at all times. Belowground clover N amounted to 40% of aboveground clover N during two consecutive years, irrespective of the nutrient availability status. Belowground clover N development was initially dominated by fast root growth, followed by enhanced root turnover during the second year. Potassium availability limited clover growth and total N accumulation in treatments with low nutrient availability. Conclusions Belowground red clover N inputs could be estimated from aboveground N by a constant factor of 0.4, regardless of the nutrient availability and cultivation time. Root turnover led to a distinct absolute increase of N rhizodeposition over time. Hence, N rhizodeposition, with an 80% share of belowground N, was the predominant N pool at the end of the second year.

Nitrogen (N) rhizodeposition is defined as the release of N from living plant roots into the soil and root turnover. The proportion of N in the soil derived from rhizodeposition (NdfR) is usually determined using 15N labelling of the plant. This isotope approach assumes that i) the enrichment of rhizodeposits is equal to the root enrichment and that ii) the root remains homogeneously enriched over space and iii) over time. The aim of this study was to quantify the bias resulting from a possible violation of the mentioned assumptions and to study the causative factors of bias. We conducted two experiments with single-pulse 15N-urea leaf-labelled red clover (Trifolium pratense L.). In the rhizodeposition experiment, we simultaneously observed the changes in substrate N concentration to obtain the mass-based rhizodeposits and determined the isotope-predicted rhizodeposits using the isotope approach. By comparing isotope-predicted to mass-based rhizodeposits we quantified the bias of the isotope approach for a period of 6 weeks. In the root distribution experiment, we observed the root 15N enrichment over space and time (4 weeks) by sampling roots grown within certain periods relative to the labelling. In both experiments we monitored the 15N distribution between shoots and roots. We observed violations of the three assumptions of the isotope approach. The average root enrichment increased over time in the root distribution experiment, but remained constant in the rhizodeposition experiment. Significant long-term translocation of 15N from shoot to root during the whole experiment (over)-compensated for growth dilution. Spatial root enrichment varied within a factor of 3, peaking in roots grown 2–8 days after labelling (d.a.l.). We observed a significant leakage of 0.5 ± 0.2% of the applied 15N within the first day after labelling corresponding to an overestimation of first-day rhizodeposits by 1100 ± 800% which translates to a calculated enrichment of 9 ± 6 atom% 15N excess for first-day rhizodeposits compared to 0.77 ± 0.09 atom% 15N excess of the root. The leaked 15N together with ordinary rhizodeposits (rhizodeposits released after the first day of labelling) led to an overestimation of N rhizodeposition by 70 ± 30% at the end of the six weeks lasting experiment using the isotope approach. The observed 15N distribution in roots and long-term 15N translocation from shoots to roots did not correspond to the expected distribution following classical pulse labelling. Thus, leaf-labelling with 15N-urea should not be considered a pure pulse-labelling method.

Root biomass is one of the most relevant root parameters for studies of plant response to environmental change, soil carbon modelling or estimations of soil carbon sequestration. A major source of error in root biomass quantification of agricultural crops in the field is the presence of extraneous organic matter in soil: dead roots from previous crops, weed roots, incorporated above ground plant residues and organic soil amendments, or remnants of soil fauna. Using the isotopic difference between recent maize root biomass and predominantly C3-derived extraneous organic matter, we determined the proportions of maize root biomass carbon of total carbon in root samples from the Swiss long-term field trial “DOK”. We additionally evaluated the effects of agricultural management (bio-organic and conventional), sampling depth (0 - 0.25, 0.25 - 0.5, 0.5 - 0.75 m) and position (within and between maize rows), and root size class (coarse and fine roots) as defined by sieve mesh size (2 and 0.5 mm) on those proportions, and quantified the success rate of manual exclusion of extraneous organic matter from root samples. Only 60 % of the root mass that we retrieved from field soil cores was actual maize root biomass from the current season. While the proportions of maize root biomass carbon were not affected by agricultural management, they increased consistently with soil depth, were higher within than between maize rows, and were higher in coarse (> 2 mm) than in fine (≤ 2 mm and > 0.5) root samples. The success rate of manual exclusion of extraneous organic matter from root samples was related to agricultural management and, at best, about 60 %. We assume that the composition of extraneous organic matter is strongly influenced by agricultural management and soil depth and governs the effect sizes of the investigated factors. Extraneous organic matter may result in severe overestimation of recovered root biomass and has, therefore, large implications for soil carbon modelling and estimations of the climate change mitigation potential of soils.